Re: brain section blowout

From:

Amos Brooks <amosbrooks@home.com>

Hi,
Sorry for the delay ...
Often it helps to try to get some sections of the block at room temp.
This will cause there to be less of a difference between the cutting temp.
and the floating temp. If you can't get a section at room temp. after the
sections are cut let them sit for a while before setting them on the float
bath to warm up in order to minimize the temp. difference.
Ya' know, it's really quite easy to make suggestions from a keyboard.
But, when the block is in front of you and giving problems, nothing seems to
work. Nonetheless, I hope this bit helps some.
good luck,
Amos Brooks
----- Original Message -----
From: "Stephanie Rodriguez" <srodriguez@primalinc.com>
To: <histonet@pathology.swmed.edu>
Sent: Friday, May 25, 2001 11:13 AM
Subject: FW: brain section blowout
> Hi,
>
> I know there was a discussion recently about tissue coming apart on the
> water bath, but I am still have trouble...
>
> I am cutting 5um sections of formalin-fixed, paraffin-embedded mouse
brain.
> When I float my ribbons out on my water bath, the sections rapidly come
> apart before I have a chance to get them on a slide.
>
> I've tried turning the temp down on my water bath, from 42C to 40C, which
> didn't change anything, then down to 35C, which, while it gave me a little
> more time, made it almost impossible to get all the wrinkles out of my
> sections, so I think that might not be warm enough!
>
> Could it be that the tissue is not adequately fixed? or perhaps poorly
> processed? I fixed the whole mouse brains in 10% NBF for 8hrs at RT, then
> 72hrs at 4C, then sent them to the contract lab that does our processing.
I
> used to work in that lab, and I know their processing schedule is pretty
> standard, but perhaps insufficient for these samples?
>
> Any suggestions would be greatly appreciated.
>
> Thanks,
>
> Stephanie Rodriguez
> Research Technologist III
> Primal, Inc.
> Seattle, WA
>
>